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(A-C) Transcriptional regulation of regenerative Ctrl and Myc +/Δ HSPCs at day 0 (D0) and day 8 (D8) post-5FU analyzed by scRNA-seq of BM LSK and LK populations: (A) scheme; (B) scFEA analysis of indicated metabolic pathways; and (C) single-cell assessment of Myc mitochondrial biogenesis score in HSC, MPP3, and GMP populations. PA, polyamine metabolism; Pyrimid, pyrimidine nucleotide biosynthesis; AA import, amino acid import. (D-E) Mitochondria quantification in regenerative Ctrl and Myc +/Δ (D) MPP3 (n=269-499 cells; 3 independent experiments) and (E) GMP (n=195-919 cells; 3 independent experiments) at D0 and D8 post-5FU analyzed by <t>Tomm20</t> immunofluorescence, with quantification of Tomm20 fluorescence (fluor.) intensity (left) and representative image (selected from 3 independent experiments; right). (F) Scheme of OXPHOS measurement by extracellular flux analysis in regenerative Ctrl and Myc +/Δ LSK at D0 and D8 post-5FU (top), with oxygen consumption rates (OCR) levels (middle) and detailed maximal respiration levels (bottom) (n=5; 4 independent experiments). O, Oligomycin A; F, FCCP; R/A, Rotenone/Antimycin A. Data are violin plots with quartiles (C) or means ± S.D. (D, E) or ± S.E.M. (F); dots represent individual cells (D, E) and circles individual mice (F); P. values were obtained by the Kruskal-Wallis test (C) or an unpaired t-test (D, E, F). See also Figures S4 and S5.
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(A-C) Transcriptional regulation of regenerative Ctrl and Myc +/Δ HSPCs at day 0 (D0) and day 8 (D8) post-5FU analyzed by scRNA-seq of BM LSK and LK populations: (A) scheme; (B) scFEA analysis of indicated metabolic pathways; and (C) single-cell assessment of Myc mitochondrial biogenesis score in HSC, MPP3, and GMP populations. PA, polyamine metabolism; Pyrimid, pyrimidine nucleotide biosynthesis; AA import, amino acid import. (D-E) Mitochondria quantification in regenerative Ctrl and Myc +/Δ (D) MPP3 (n=269-499 cells; 3 independent experiments) and (E) GMP (n=195-919 cells; 3 independent experiments) at D0 and D8 post-5FU analyzed by <t>Tomm20</t> immunofluorescence, with quantification of Tomm20 fluorescence (fluor.) intensity (left) and representative image (selected from 3 independent experiments; right). (F) Scheme of OXPHOS measurement by extracellular flux analysis in regenerative Ctrl and Myc +/Δ LSK at D0 and D8 post-5FU (top), with oxygen consumption rates (OCR) levels (middle) and detailed maximal respiration levels (bottom) (n=5; 4 independent experiments). O, Oligomycin A; F, FCCP; R/A, Rotenone/Antimycin A. Data are violin plots with quartiles (C) or means ± S.D. (D, E) or ± S.E.M. (F); dots represent individual cells (D, E) and circles individual mice (F); P. values were obtained by the Kruskal-Wallis test (C) or an unpaired t-test (D, E, F). See also Figures S4 and S5.
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(A-C) Transcriptional regulation of regenerative Ctrl and Myc +/Δ HSPCs at day 0 (D0) and day 8 (D8) post-5FU analyzed by scRNA-seq of BM LSK and LK populations: (A) scheme; (B) scFEA analysis of indicated metabolic pathways; and (C) single-cell assessment of Myc mitochondrial biogenesis score in HSC, MPP3, and GMP populations. PA, polyamine metabolism; Pyrimid, pyrimidine nucleotide biosynthesis; AA import, amino acid import. (D-E) Mitochondria quantification in regenerative Ctrl and Myc +/Δ (D) MPP3 (n=269-499 cells; 3 independent experiments) and (E) GMP (n=195-919 cells; 3 independent experiments) at D0 and D8 post-5FU analyzed by <t>Tomm20</t> immunofluorescence, with quantification of Tomm20 fluorescence (fluor.) intensity (left) and representative image (selected from 3 independent experiments; right). (F) Scheme of OXPHOS measurement by extracellular flux analysis in regenerative Ctrl and Myc +/Δ LSK at D0 and D8 post-5FU (top), with oxygen consumption rates (OCR) levels (middle) and detailed maximal respiration levels (bottom) (n=5; 4 independent experiments). O, Oligomycin A; F, FCCP; R/A, Rotenone/Antimycin A. Data are violin plots with quartiles (C) or means ± S.D. (D, E) or ± S.E.M. (F); dots represent individual cells (D, E) and circles individual mice (F); P. values were obtained by the Kruskal-Wallis test (C) or an unpaired t-test (D, E, F). See also Figures S4 and S5.
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(A-C) Transcriptional regulation of regenerative Ctrl and Myc +/Δ HSPCs at day 0 (D0) and day 8 (D8) post-5FU analyzed by scRNA-seq of BM LSK and LK populations: (A) scheme; (B) scFEA analysis of indicated metabolic pathways; and (C) single-cell assessment of Myc mitochondrial biogenesis score in HSC, MPP3, and GMP populations. PA, polyamine metabolism; Pyrimid, pyrimidine nucleotide biosynthesis; AA import, amino acid import. (D-E) Mitochondria quantification in regenerative Ctrl and Myc +/Δ (D) MPP3 (n=269-499 cells; 3 independent experiments) and (E) GMP (n=195-919 cells; 3 independent experiments) at D0 and D8 post-5FU analyzed by <t>Tomm20</t> immunofluorescence, with quantification of Tomm20 fluorescence (fluor.) intensity (left) and representative image (selected from 3 independent experiments; right). (F) Scheme of OXPHOS measurement by extracellular flux analysis in regenerative Ctrl and Myc +/Δ LSK at D0 and D8 post-5FU (top), with oxygen consumption rates (OCR) levels (middle) and detailed maximal respiration levels (bottom) (n=5; 4 independent experiments). O, Oligomycin A; F, FCCP; R/A, Rotenone/Antimycin A. Data are violin plots with quartiles (C) or means ± S.D. (D, E) or ± S.E.M. (F); dots represent individual cells (D, E) and circles individual mice (F); P. values were obtained by the Kruskal-Wallis test (C) or an unpaired t-test (D, E, F). See also Figures S4 and S5.
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(A-C) Transcriptional regulation of regenerative Ctrl and Myc +/Δ HSPCs at day 0 (D0) and day 8 (D8) post-5FU analyzed by scRNA-seq of BM LSK and LK populations: (A) scheme; (B) scFEA analysis of indicated metabolic pathways; and (C) single-cell assessment of Myc mitochondrial biogenesis score in HSC, MPP3, and GMP populations. PA, polyamine metabolism; Pyrimid, pyrimidine nucleotide biosynthesis; AA import, amino acid import. (D-E) Mitochondria quantification in regenerative Ctrl and Myc +/Δ (D) MPP3 (n=269-499 cells; 3 independent experiments) and (E) GMP (n=195-919 cells; 3 independent experiments) at D0 and D8 post-5FU analyzed by Tomm20 immunofluorescence, with quantification of Tomm20 fluorescence (fluor.) intensity (left) and representative image (selected from 3 independent experiments; right). (F) Scheme of OXPHOS measurement by extracellular flux analysis in regenerative Ctrl and Myc +/Δ LSK at D0 and D8 post-5FU (top), with oxygen consumption rates (OCR) levels (middle) and detailed maximal respiration levels (bottom) (n=5; 4 independent experiments). O, Oligomycin A; F, FCCP; R/A, Rotenone/Antimycin A. Data are violin plots with quartiles (C) or means ± S.D. (D, E) or ± S.E.M. (F); dots represent individual cells (D, E) and circles individual mice (F); P. values were obtained by the Kruskal-Wallis test (C) or an unpaired t-test (D, E, F). See also Figures S4 and S5.

Journal: bioRxiv

Article Title: Glutamine addiction is a therapeutic target to block emergency myelopoiesis

doi: 10.64898/2026.03.26.714544

Figure Lengend Snippet: (A-C) Transcriptional regulation of regenerative Ctrl and Myc +/Δ HSPCs at day 0 (D0) and day 8 (D8) post-5FU analyzed by scRNA-seq of BM LSK and LK populations: (A) scheme; (B) scFEA analysis of indicated metabolic pathways; and (C) single-cell assessment of Myc mitochondrial biogenesis score in HSC, MPP3, and GMP populations. PA, polyamine metabolism; Pyrimid, pyrimidine nucleotide biosynthesis; AA import, amino acid import. (D-E) Mitochondria quantification in regenerative Ctrl and Myc +/Δ (D) MPP3 (n=269-499 cells; 3 independent experiments) and (E) GMP (n=195-919 cells; 3 independent experiments) at D0 and D8 post-5FU analyzed by Tomm20 immunofluorescence, with quantification of Tomm20 fluorescence (fluor.) intensity (left) and representative image (selected from 3 independent experiments; right). (F) Scheme of OXPHOS measurement by extracellular flux analysis in regenerative Ctrl and Myc +/Δ LSK at D0 and D8 post-5FU (top), with oxygen consumption rates (OCR) levels (middle) and detailed maximal respiration levels (bottom) (n=5; 4 independent experiments). O, Oligomycin A; F, FCCP; R/A, Rotenone/Antimycin A. Data are violin plots with quartiles (C) or means ± S.D. (D, E) or ± S.E.M. (F); dots represent individual cells (D, E) and circles individual mice (F); P. values were obtained by the Kruskal-Wallis test (C) or an unpaired t-test (D, E, F). See also Figures S4 and S5.

Article Snippet: Slides were stained with rabbit anti-Tomm20 primary antibody (Cell Signaling Technology, Cat#42406; 1:100) in Block/Perm media for 2 hours at RT, washed, and stained with donkey anti-rabbit IgG AF488 secondary antibody (Jackson ImmunoResearch, Cat#711-545-152; 1:400) and DAPI (5 μg/ml) for 1 hour at RT.

Techniques: Single Cell, Immunofluorescence, Fluorescence